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Infection and Immunity, October 2005, p. 6812-6821, Vol. 73, No. 10
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.10.6812-6821.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Successful Selection of Cross-Protective Vaccine Candidates for Ornithobacterium rhinotracheale Infection

D. F. Schuijffel,1 P. C. M. van Empel,1 A. M. M. A. Pennings,1 J. P. M. van Putten,2 and P. J. M. Nuijten3*

Intervet International BV, Bacteriology R&D, Boxmeer, The Netherlands,1 Utrecht University, Faculty of Veterinary Medicine, Department of Infectious Diseases and Immunology, Utrecht, The Netherlands,2 Nobilon International BV, Boxmeer, The Netherlands3

Received 6 September 2004/ Returned for modification 14 December 2004/ Accepted 31 May 2005

Ornithobacterium rhinotracheale is a bacterial pathogen known for causing respiratory disease in poultry. In this study, we demonstrate for the first time that cross-protective immunity against different O. rhinotracheale serotypes can be induced by live vaccination. Sera from these live-vaccinated and cross-protected birds were used to identify new vaccine targets by screening an O. rhinotracheale expression library. Out of 20,000 screened plaques, a total of 30 cross-reactive clones were selected for further analysis. Western blot analysis and DNA sequencing identified eight different open reading frames. The genes encoding the eight cross-reactive antigens were amplified, cloned in an expression vector, and expressed in Escherichia coli. Purified recombinant proteins with a molecular mass ranging from 35.9 kDa to 62.9 kDa were mixed and tested as a subunit vaccine for (cross-)protection against challenge with homologous and heterologous O. rhinotracheale serotypes in chickens. Subunit vaccination resulted in the production of antibodies reactive to the recombinant proteins on Western blot, and this eight-valent vaccine conferred both homologous and heterologous protection against O. rhinotracheale challenge in chickens.

* Corresponding author. Mailing address: Bacteriological R&D, Nobilon International BV, P.O. Box 320, Exportstraat 39b, 5830 AH Boxmeer, The Netherlands. Phone: 31 (0) 485 58 7326. Fax: 31 (0) 485 58 7490. E-mail: Piet.Nuijten{at}Nobilonvaccines.com.

Editor: J. F. Urban, Jr.

Infection and Immunity, October 2005, p. 6812-6821, Vol. 73, No. 10
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.10.6812-6821.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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Copyright © 2005 by the American Society for Microbiology. All rights reserved.